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Highly specific capture and direct MALDI-MS analysis of phosphorylated peptides using novel multifunctional chitosan-GMA-IDA-Fe (III) nanosphere

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Abstract

In this study, we describe a method for highly specific enrichment of phosphopeptides with multifunctional chitosan–glycidyl methacrylate (GMA)–iminodiacetic acid (IDA)–Fe (III) nanospheres for direct analysis by matrix-assisted laser desorption–ionization mass spectrometry (MALDI-MS). This is the first time that chitosan has been used to create nanospheres support material for selective enrichment of phosphopeptides by modification with GMA, derivatization with IDA, and loading with Fe (III) ions. Chitosan-GMA-IDA-Fe (III) nanospheres with a diameter of 20 to 100 nm have multifunctional chemical moieties which confer unique properties, good dispersibility in highly acidic binding buffers, as well as good biocompatibility and chemical stability which improves their specific interaction with phosphopeptides using various types of acid binding buffers. The process of enrichment is very simple, quick, efficient, and specific. Its high specificity and efficiency for purification of phosphopeptides is reflected in the very low and substoichiometric amounts of phosphopeptides which can be detected, in quantities as low as 1:3,000 M ratios. Compared with other state-of the-art technologies such as the use of conventional Fe3+-IMAC and TiO2, these chitosan nanosphere techniques show superior specificity and sensitivity. Moreover, the resultant chitosan-GMA-IDA-Fe3+ nanosphere-absorbed phosphopeptides can be either directly analyzed by MALDI-TOF MS analysis or eluted and further analyzed by nano-LC-MS/MS.

A method for highly specific enrichment of phosphopeptides with novel multifunctional chitosan-GMA-IDA-Fe (III) nanospheres for direct analysis by matrix assisted laser desorption-ionization mass spectrometry (MALDI MS).

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Acknowledgement

This work was supported by the “985 project” of China. We thank Shimadzu Group Company for supporting the MALDI-TOF MS.

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Authors and Affiliations

  1. Proteomics Laboratory, Medical and Healthy Analytical Center, Peking University, Beijing, 100191, China

    Xiajuan Zou, Dan Liu, Lijun Zhong, Bin Yang, Yaxin Lou & Yuxin Yin

  2. Microscope Electronic Laboratory, Medical and Healthy Analytical Center, Peking University, Beijing, 100191, China

    Baihe Hu

  3. Institute of Systems Biomedicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100191, China

    Yuxin Yin

Authors
  1. Xiajuan Zou
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  2. Dan Liu
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  3. Lijun Zhong
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  4. Bin Yang
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  5. Yaxin Lou
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  6. Baihe Hu
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Correspondence to Xiajuan Zou or Yuxin Yin.

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Zou, X., Liu, D., Zhong, L. et al. Highly specific capture and direct MALDI-MS analysis of phosphorylated peptides using novel multifunctional chitosan-GMA-IDA-Fe (III) nanosphere. Anal Bioanal Chem 401, 1251–1261 (2011). https://doi.org/10.1007/s00216-011-5186-x

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  • DOI: https://doi.org/10.1007/s00216-011-5186-x

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